Rapid diagnostics of genital herpes by loop-mediated isothermal amplification method with fluorescent detection

Смирнова, Д. И.; Петруша, О. А.; Грачёва, А. В.; Волынская, Е. А.; Зверев, В. В.; Файзулоев, Е. Б.

doi:10.36233/0372-9311-2019-6-40-46

Cited by 2 publications

(1 citation statement)

References 7 publications

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“…Loop-Mediated Isothermal Amplification Technique: DNA amplification, with high efficiency and specificity, can also be achieved using the loop-mediated isothermal amplification (LAMP) method [ 28 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 ]. It involves the use of Bst DNA polymerase and a set of six primers that recognize specific sequences on the target DNA.…”

Section: Detection Of Hsv In Lesionsmentioning

confidence: 99%

Diagnosis of Herpes Simplex Virus: Laboratory and Point-of-Care Techniques

Nath

Kabir

Doust

et al. 2021

Infectious Disease Reports

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Herpes is a widespread viral infection caused by the herpes simplex virus (HSV) that has no permanent cure to date. There are two subtypes, HSV-1 and HSV-2, that are known to cause a variety of symptoms, ranging from acute to chronic. HSV is highly contagious and can be transmitted via any type of physical contact. Additionally, viral shedding can also happen from asymptomatic infections. Thus, early and accurate detection of HSV is needed to prevent the transmission of this infection. Herpes can be diagnosed in two ways, by either detecting the presence of the virus in lesions or the antibodies in the blood. Different detection techniques are available based on both laboratory and point of care (POC) devices. Laboratory techniques include different biochemical assays, microscopy, and nucleic acid amplification. In contrast, POC techniques include microfluidics-based tests that enable on-spot testing. Here, we aim to review the different diagnostic techniques, both laboratory-based and POC, their limits of detection, sensitivity, and specificity, as well as their advantages and disadvantages.

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Section: Detection Of Hsv In Lesionsmentioning

confidence: 99%

Diagnosis of Herpes Simplex Virus: Laboratory and Point-of-Care Techniques

Nath

Kabir

Doust

et al. 2021

Infectious Disease Reports

View full text Add to dashboard Cite

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Diagnostic value of the LAMP method with real-time fluo-rescence detection on a model of herpesvirus infection

Смирнова

Gracheva

Volynskaya

et al. 2021

Sanitarnyj Vrač (Sanitary Doctor)

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Due to the high prevalence and clinical significance of herpesvirus diseases, improvement of methods of their diagnostic remains relevant.The aim of the work: improvement of the methodological approaches to the rapid differential diagnosis of herpesvirus infections based on the LAMP method with real-time fluorescence detection (LAMPRT). 201 urogenital swabs were examined using RT-PCR and LAMP-RT, 27of which contained DNAof Epstein-Barr virus (EBV), 34 — Cytomegalovirus (CMV), 14 — Herpes simplex virus type 1 (HSV-1), 36 — Herpes simplex virus type 2 (HSV-2). For LAMP-RT reaction we used Bst 2.0 WarmStart DNA polymerase (BioLabs, Great Britain), SYTO-82 dye (Invitrogen, USA), and sets of primers for herpesvirus DNA detection by LAMP. High efficiency of using the SYTO-82 dye for the detection of herpesvirus DNA in the LAMP-RT reactionwas shown. Under the optimal conditions, the LAMP-RT allows to reduce the reaction time to 25 minutes, that 3 times less then real-time PCR. The diagnostic sensitivity of the LAMP-RT reaction for HSV-1 was 100 %, HSV-2 — 94.50 %, EBV — 89 %, CMV — 94 %. The diagnostic specificity for all studied viruses was 100 %. The analytical sensitivity of EBV detection was 104 DNA copies/ ml, for HSV types 1 and 2 and CMV — 103 DNA copies/ml. Thus, the LAMP-RT reaction with SYTO-82 dye makes it possible to detect the DNA of various herpesviruses in clinical specimens with high sensitivity and specificity and can be considered as a promising method for point-of-care diagnostics. For the widespread implementation of the method into the practice of laboratory diagnostics, it is necessary to solve the problem of creating an internal positive control of the reaction and the development of portable specialized analyzers.

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Rapid diagnostics of genital herpes by loop-mediated isothermal amplification method with fluorescent detection

Cited by 2 publications

References 7 publications

Diagnosis of Herpes Simplex Virus: Laboratory and Point-of-Care Techniques

Diagnosis of Herpes Simplex Virus: Laboratory and Point-of-Care Techniques

Diagnostic value of the LAMP method with real-time fluo-rescence detection on a model of herpesvirus infection

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